Canine Bile Research Articles

Feasibility and safety of trans-biliary cryoablation: Preclinical evaluation of a novel flexible cryoprobe

Cryoablation, as a well-characterized technology, has multifarious clinical applications in solid malignancy. However, trans-biliary cryoablation for malignant biliary obstruction has not been reported yet. Thus, this study aimed to determine the efficacy and safety of trans-biliary cryoablation with a novel CO2 gas-based flexible cryoprobe in standardized preclinical settings. For fresh porcine liver ex vivo, the freezing efficacy of cryoablation was evaluated by using fresh porcine liver. The real-time CO2 flow rate, freezing temperature and freezing range were examined and the frozen appearance was visualized. In vivo study, acute and chronical effects were investigated by using the models of canine bile duct. Histopathology and laboratory examination were performed. The lowest temperature that the electrode could deliver to the tissue was −60.7 °C. At 60s after freezing, the tissue temperature dropped to −22.6 °C and −4.3 °C at 0.1 and 0.2 cm from the electrode center, respectively. The frozen size was greater in liver tissue ex vivo than that in bile duct tissue in vivo. No biliary hemorrhage, perforation, stricture, obstruction, and adjacent organ injury were observed. With histopathologic examination, acute intercellular vacuoles were observed in the lamina propria adjacent to the lumen. Chronic changes, including uneven coagulative necrosis, fibro-proliferation, inflammatory infiltration and connective tissue thickening were observed in the lamina propria of the all biliary samples. The results demonstrated CO2 gas-based trans-biliary cryoablation is safe and efficacious. These findings may provide a potential new modality for primary malignant biliary obstruction and malignant obstruction within a biliary stent and contribute to cryoablation of clinical practice.

In vitro effect of 20% N-acetylcysteine on the viscosity of normal canine bile.

OBJECTIVE To evaluate the in vitro effect of 20% N-acetylcysteine (NAC) on the viscosity of normal canine bile. ANIMALS Bile samples obtained from 10 adult dogs euthanized for reasons unrelated to biliary disease. PROCEDURES Each sample was centrifuged to remove particulates, then divided into 3 aliquots. One aliquot remained untreated (control). Each of the other aliquots was diluted 1:4 with 20% NAC or sterile water. The viscosity of all samples was measured with a rotational viscometer at 25°C. Viscosity of control samples was measured immediately after centrifugation and at 1 and 24 hours after treatment application to the diluted samples. Viscosity of diluted samples was measured at 1 and 24 hours after treatment application. RESULTS Mean viscosity differed significantly among the 3 groups at both 1 and 24 hours after treatment application. Relative to control samples, the addition of NAC and sterile water decreased the viscosity by approximately 3.35 mPa·s (95% confidence interval [CI], 1.58 to 5.12 mPa·s) and 2.74 mPa·s (95% CI, 1.33 to 4.14 mPa·s), respectively. Mean viscosity of the NAC-treated samples was approximately 0.61 mPa·s (95% CI, 0.21 to 1.01 mPa·s) less than that for the sterile water-treated samples. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that in vitro dilution of canine bile 1:4 with 20% NAC significantly decreased the viscosity of the resulting mixture. Further research is necessary to determine whether NAC is a safe and effective noninvasive treatment for dogs with persistent biliary sludge or gallbladder mucoceles.

Erythrocyte copper chaperone for superoxide dismutase and superoxide dismutase as biomarkers for hepatic copper concentrations in Labrador retrievers

Hereditary hepatic copper accumulation in Labrador retrievers leads to hepatitis with fibrosis and eventually cirrhosis. The development of a non-invasive blood-based biomarker for copper status in dogs could be helpful in identifying dogs at risk and to monitor copper concentrations during treatment. In this study, two cellular copper metabolism proteins, Cu/Zn superoxide dismutase (SOD1) and its chaperone (copper chaperone for SOD1, CCS) were measured in erythrocytes and tested for association with hepatic copper concentrations in 15 Labrador retrievers with normal or increased hepatic copper concentrations.Antibodies against CCS and SOD1 were applicable for use in canine specimens. This was demonstrated by the loss of immune-reactive bands for CCS and SOD1 in siRNA treated canine bile duct epithelial cells. Erythrocyte CCS and CCS/SOD1 ratios were decreased 2.37 (P < 0.001) and 3.29 (P < 0.001) fold in the high copper group compared to the normal copper group. Erythrocyte CCS and CCS/SOD1 ratio are potential new biomarkers for hepatic copper concentrations in Labrador retrievers and could facilitate early diagnosis and treatment monitoring for copper-associated hepatitis in dogs.

Evaluation of a semiquantitative SNAP test for measurement of bile acids in dogs.

Background. Serum bile acids (SBA) are used as a routine screening tool of liver function in dogs. Serum samples are usually shipped to a referral laboratory for quantitative analysis with an enzymatic chemistry analyzer. The canine SNAP Bile Acids Test (SNAP-BAT) provides an immediate, semi-quantitative measurement of bile acid concentrations in-house. With the SNAP-BAT, bile acids concentrations of 5–30 µmol/L are quantified, and results outside of that range are classified as <5 or >30 µmol/L. Agreement of the SNAP-BAT with the enzymatic method has not been extensively investigated.Objectives. The purposes of this prospective clinical study were to assess the precision of the SNAP-BAT and determine agreement of SNAP-BAT with results from an in-house chemistry analyzer.Methods. After verifying intra-assay precision of the SNAP-BAT, a prospective analysis was performed using blood samples collected from 56 dogs suspected to have liver disease. Each sample was analyzed with an enzymatic, in-house chemistry analyzer and the SNAP-BAT. Agreement between the two methods was statistically assessed using the κ index of agreement.Results. Intra-assay variability was minimal. The κ index for agreement between the SNAP-BAT and routine chemistry analyzer was between 0.752 and 0.819, indicating substantial to near perfect agreement.Conclusions. The SNAP-BAT is a highly accurate, semi-quantitative test that yields immediate results, and has very little intra-assay variability, particularly for results >30 µmol/L.

Cefotaxime-Eluting Covered Self-Expandable Stents in a Canine Biliary Model: Scanning Electron Microscopic Study of Biofilm Formation

As bacterial attachment and subsequent biofilm formation on the surface of a stent lumen may be important in initiating sludge formation and subsequent stent blockage, there is a need for antibiotic-eluting stents to prevent sludge formation by bacterial attachment and colonization. To evaluate the antimicrobial effects of cefotaxime-eluting covered self-expandable stents (CE-CSEMS) in a canine biliary model by using scanning electron microscopy (SEM) to analyze biofilm formation on the inner surface of the stents. Three 10% (wt/vol) CE-CSEMS, three 20% (wt/vol) CE-CSEMS, and three control stents (CS) were endoscopically inserted into the bile ducts of nine mongrel dogs, which were followed up for 8 weeks. Gross and SEM examinations were performed to access the development and degree of biofilm formation in dogs with CS and CE-CSEMS. All stents were successfully inserted into canine bile ducts without any procedure-related complications. All nine dogs remained in good health without jaundice over the next 8 weeks. The degree of biofilm was similar in dogs implanted with CS and CE-CSEMS using two concentrations of cefotaxime either by gross inspection or SEM. All stent types showed a similar biofilm structure with relatively uniform thickness on SEM. Cefotaxime does not prevent biofilm development in a canine biliary model. Biofilm formation on the inner surface of biliary stents may be prevented by using broad-spectrum antimicrobial agents and stents that maintain a steady release of these agents for a long time, thus prolonging stent patency.

Expanded polytetrafluoroethylene in canine bile duct injury: a critical analysis

Analyze the morphological and structural outcomes of a patch of expanded polytetrafluoroethylene in the treatment of an iatrogenic injury of the common bile duct. In Group 1 (Sham), 7 dogs underwent 3 laparotomies with intervals of 30 days between them. In Group 2, 10 dogs underwent transient common bile duct obstruction. After 30 days, this biliary occlusion was undone and a patch of expanded polytetrafluoroethylene replaced a fragment removed from the duct's wall. Thirty days after this last surgery, cholangiographic assessment of prosthesis patency and macro and microscopic evaluation of the biliary tract were performed. Daily clinical inspection completed the study outcomes. The Wilcoxon non-parametric test was used for statistical analysis. In all dogs enlargement of the biliary tree diameter was observed 30 and 60 days after the first surgical procedure. Partial adhesion of the patch to the common bile duct as a free luminal foreign body was found in 6 dogs. The prosthesis was completely integrated to surrounding tissue in the remaining four. Although a feasible option for the treatment of biliary duct iatrogenic lesions, the expanded polytetrafluoroethylene prosthesis must be used with caution considering the potential risks for complications.

Experimental Study of Poly-l-Lactic Acid Biodegradable Stents in Normal Canine Bile Ducts

This study was designed to clarify the advantages of biodegradable stents in terms of mucosal reaction and biodegradation after placement. We designed a biodegradable stent and assessed stent degradation and changes in the normal bile ducts of dogs. The biodegradable stent is a balloon-expandable Z stent consisting of poly-L-lactic acid (PLLA) with a diameter of 6 mm and a length of 15 mm. We assessed four groups of three beagle dogs each at 1, 3, 6, and 9 months of follow-up. After evaluating stent migration by radiography and stent and bile duct patency by cholangiography, the dogs were sacrificed to remove the bile duct together with the stent. The bile duct lumen was examined macroscopically and histologically, and the stent degradation was examined macroscopically and by scanning electron microscopy (SEM). Bile duct obstruction was absent and none of the stents migrated. Macroscopic evaluation showed moderate endothelial proliferation in the bile ducts at the implant sites at 3 and 6 months and a slight change at 9 months. Slight mononuclear cell infiltration was histologically identified at all time points and epithelial hyperplasia that was moderate at 3 months was reduced to slight at 6 and 9 months. Stent degradation was macroscopically evident in all animals at 9 months and was proven by SEM in two dogs at 6 months and in all of them at 9 months. Our results suggest that PLLA bioabsorbable stents seems to be useful for implantation in the biliary system with further investigation.

Histologic influence of paclitaxel-eluting covered metallic stents in a canine biliary model

Metallic stents for malignant biliary obstruction are susceptible to occlusion by tumor ingrowth or overgrowth. Paclitaxel-eluting metallic stents (PEMSs), however, may prevent occlusion from tumor ingrowth. We evaluated the safety of PEMSs in a canine biliary model by analyzing tissue reactions by histologic examination. Animal study. Six PEMSs and 5 control stents (CSs) were endoscopically placed in the bile ducts of 11 mongrel dogs and followed up for 6 weeks. The animals were euthanized at 6 weeks, and gross and microhistologic examinations were done. A comparison was made between dogs with PEMSs and dogs with CSs. All 11 stents were successfully placed in the bile ducts, although the distal ends of 2 PEMSs were partially exposed to the duodenal lumen. All 11 dogs survived until death without evidence of jaundice. Gross histologic examination showed mucosal hyperplasia in 3 of 6 dogs in the PEMS group, but there was no evidence of mucosal hyperplasia in the CS group. Microscopically, the epithelial layers at the middle and distal ends of the stented segments were significantly thicker in the PEMS dogs than in the CS group. Placement of PEMSs in normal canine bile ducts was without significant technical complications; however, local drug delivery from the PEMS resulted in significant histologic changes. Therefore, it is important to choose a drug in adequate concentrations to exert an antitumor effect without damaging the adjacent normal bilary mucosa.

Influence of a Paclitaxel-Eluting Covered Metallic Stent On Tissue Hyperplasia: An Experimental Study in a Canine Biliary Model

Aim: The aim was to evaluate safety of a paclitaxel-eluting covered metallic stent in a canine biliary model by comparing histologic tissue reaction. Methods: Six paclitaxel-eluting covered metallic stents were endoscopically placed in the bile ducts of six mongrel dogs, and five covered metallic stents were placed in five dogs as controls. The dogs were sacrificed at six weeks after stent placement and gross and histopathologic examinations were done. Degree of inflammation was graded as follows: 0, none; 1, mild; 2, moderate; 3, severe. Results: All 11 dogs survived until the sacrifice without evidence of jaundice. All the stents except for two drug-eluting stents were placed within the canine bile duct successfully. Two malpositioned stents were excluded from histologic analysis. There was no migration or perforation. In the control stent group, there was no tissue ingrowth. Both proximal and distal ends of all the control stents showed minimal tissue overgrowth except for distal end obstruction in one and 20% narrowing of proximal end in another. In the drug-eluting stent group, tissue ingrowth was found in three animals. Two were malpositioned stents. Minimal tissue overgrowth was noted in both ends of three animals. One dog without malpositioned stent showed ingrowth at both ends. Microscopic examination demonstrated increased thickness of epithelial layers in middle and distal ends of canine bile ducts in contact with drug-eluting stents (575.00 ± 301.39 μm vs. 180.00 ± 109.55 μm, P = 0.022; 1150.00 ± 100.00 μm vs. 430.00 ± 323.27 μm, P = 0.012, respectively). Proximal ends of bile ducts showed no significant difference in thickness of epithelial layers, but there was tendency for increased thickness in the drug-eluting stent group (800.00 ± 282.84 μm vs. 530.00 ± 272.95 μm, P = 0.260). Canine bile duct in contact with drug-eluting stents showed less severe inflammatory reaction in the proximal and distal ends (2 vs. 2.80 ± 0.45, P = 0.024; 1.50 ± 0.58 vs. 2.60 ± 0.55, P = 0.036, respectively). In the middle of canine bile duct in contact with drug-eluting stents, there was tendency for less severe inflammation although not statistically significant (1.50 ± 1.00 vs. 1.80 ± 0.84, P = 0.498). Conclusion: Placement of paclitaxel-eluting covered metallic stents in normal canine bile duct was without significant complications despite significant histologic changes from local drug delivery. The results provide a basis for extending the use of a paclitaxel-eluting covered metallic stent in clinical trials. Further studies in patients with malignant biliary obstruction are needed to establish the role of this stent in the palliation of obstructive jaundice.

Canine Intestinal Contents vs. Simulated Media for the Assessment of Solubility of Two Weak Bases in the Human Small Intestinal Contents

This study was conducted to assess the relative usefulness of canine intestinal contents and simulated media in the prediction of solubility of two weak bases (dipyridamole and ketoconazole) in fasted and fed human intestinal aspirates that were collected under conditions simulating those in bioavailability/bioequivalence studies. After administration of 250 mL of water or 500 mL of Ensure plus [both containing 10 mg/mL polyethylene glycol (PEG) 4000 as nonabsorbable marker], intestinal aspirates were collected from the fourth part of the duodenum of 12 healthy adults and from the mid-jejunum of four Labradors. Pooled samples were analyzed for PEG, pH, buffer capacity, osmolality, surface tension, pepsin, total carbohydrates, total protein content, bile salts, phospholipids, and neutral lipids. The shake-flask method was used to measure the solubility of dipyridamole and ketoconazole in pooled human and canine intestinal contents and in fasted-state-simulating intestinal fluid (FaSSIF) and fed-state-simulating intestinal fluid (FeSSIF) containing various bile salts and pH-buffering agents. For both compounds, solubility in canine contents may be predictive of human intralumenal solubility in the fasting state but not in the fed state. The poor agreement of results in canine and human aspirates can be attributed to the higher bile salt content in canine bile. Solubility in FaSSIF containing a mixture of bile salts from crude bile predicted satisfactorily the intralumenal solubility of both drugs in the fasted state in humans. Solubility in FeSSIF, regardless of the identity of bile salts or of the buffering species, deviated from intralumenal values in the fed human aspirates by up to 40%. This was attributed to the lack of lipolytic products in FeSSIF, the higher bile salt content of FeSSIF, and the lower pH of FeSSIF. FaSSIF containing a mixture of bile salts from crude bile, and FeSSIF containing lipolytic products and, perhaps, having lower bile salt content but slightly higher pH, should be more useful than canine intestinal aspirates for predicting intralumenal solubilities in humans.

Esophageal ATP synthase and keratinocyte growth factor gene expression changes after acid and bile-induced mucosal damage

Intramural gene expression changes may be critically involved in tissue damage, defense and repair after esophageal regurgitation. The aims were to characterize the consequences of short-term exposure to luminal bile, acid, or bile mixed with acid on the beta-ATPase, keratinocyte growth factor 1 (KGF-1) and KGF receptor (KGF-R) expressions within the mucosa and the muscle layer in a large animal model. Esophageal segments of anesthetized dogs were exposed to saline (n = 3), diluted canine bile (n = 6), hydrochloric acid (n = 5) or bile + hydrochloric acid (n = 5), and tissue biopsies were taken at the end of the 180-min observation period. Semiquantitative reverse transcriptase polymerase chain reactions were carried out and the degree of histological damage was evaluated on the 0-16-grade Geisinger scoring scale. Acid exposure was followed by a significant decrease in the level of beta-ATPase expression in the mucosa, and parallel increases in KGF-1 and KGF-R expression. Corresponding changes in the muscle layer were not significant. Bile alone evoked more severe tissue damage, with significantly decreased beta-ATPase levels in both the mucosa and the muscle, whereas the KGF-1 expression did not change significantly. The bile + acid treatment induced an intermediate state, with significant beta-ATPase transcription level decreases in both layers, while the mucosal KGF-1 expression was lower than that following acid treatment alone. The acid-induced transcriptional level downregulation of mucosal beta-ATPase gene expression in the smooth muscle layer was exacerbated by bile, but the concomitant KGF and KGF-R gene expression changes may indicate the start of a consecutive repair process.

Bile-induced adenosine triphosphate depletion and mucosal damage during reflux esophagitis.

This study was designed to investigate the role of bile in a large animal model of acute esophageal reflux disease. An agar electrode was used to measure the transmucosal potential difference of the esophagus in anaesthetized dogs. The vascular permeability index and the epithelial permeability index of the mucosa were evaluated by means of the Evans blue and the sodium-fluorescein clearance method, respectively. The tissue adenosine triphosphate (ATP) level and the myeloperoxidase activity were determined from tissue biopsies, while the degree of mucosal damage was evaluated histologically on a grade 0-100 scale. Group 1 (n = 8) served as saline-treated control; groups 2 (n = 8), 3 (n = 5) and 4 (n = 5) were exposed for 4 h to canine bile alone, to hydrochloric acid + bile, or to hydrochloric acid alone, respectively. In Groups 2, 3 and 4 the degree of mucosal damage was significantly increased, and a 4-fold elevation in myeloperoxidase activity was observed. The transmucosal potential difference was decreased significantly below the control level, while the vascular and epithelial permeability indices were significantly increased compared with the control values. Bile, but not hydrochloric acid, evoked a significant (40%) decrease in the ATP level of the esophageal tissue. We propose that mucosal dysfunction, structural damage and leukocyte invasion during hydrochloric acid-induced esophageal injury are exacerbated by bile-induced changes in tissue ATP concentrations during experimental esophageal reflux disease.

Bile-induced Adenosine Triphosphate Depletion and Mucosal Damage During Reflux Esophagitis

Background: This study was designed to investigate the role of bile in a large animal model of acute esophageal reflux disease. Methods: An agar electrode was used to measure the transmucosal potential difference of the esophagus in anaesthetized dogs. The vascular permeability index and the epithelial permeability index of the mucosa were evaluated by means of the Evans blue and the sodium-fluorescein clearance method, respectively. The tissue adenosine triphosphate (ATP) level and the myeloperoxidase activity were determined from tissue biopsies, while the degree of mucosal damage was evaluated histologically on a grade 0-100 scale. Group 1 (n = 8) served as saline-treated control; groups 2 (n = 8), 3 (n = 5) and 4 (n = 5) were exposed for 4 h to canine bile alone, to hydrochloric acid + bile, or to hydrochloric acid alone, respectively. Results: In Groups 2, 3 and 4 the degree of mucosal damage was significantly increased, and a 4-fold elevation in myeloperoxidase activity was observed. The transmucosal potential difference was decreased significantly below the control level, while the vascular and epithelial permeability indices were significantly increased compared with the control values. Bile, but not hydrochloric acid, evoked a significant (40%) decrease in the ATP level of the esophageal tissue. Conclusions: We propose that mucosal dysfunction, structural damage and leukocyte invasion during hydrochloric acid-induced esophageal injury are exacerbated by bile-induced changes in tissue ATP concentrations during experimental esophageal reflux disease.

The effect of prostanoids on hepatic bile flow in dogs with normal liver and bile duct cell hyperplasia

Bile flow rates and composition are subject to a wide variety of neural, endocrine and paracrine influences. The effects of these multiple factors may be different in the diseased liver compared to the response produced in the normal liver. As prostanoids may have a therapeutic role in liver disease it was intended to evaluate the effects of two principal therapeutic prostanoids, prostaglandin E 2 and prostacyclin, on bile flow in dogs with a normal liver and in dogs with hepatotoxin-induced liver injury. Initially, in awake animals with chronic biliary and gastric fistulas the bile flow response to prostaglandin E 2 and prostacyclin was evaluated and compared to the response produced by bile salt infusion alone and to that produced by the standard choleretic hormones, secretin and glucagon. The animals were then fed α-naphthylisothiocyanate (ANIT) and the studies repeated. ANIT is a hepatotoxin that produces bile duct cell hyperplasia which was confirmed in dogs by demonstrating that ANIT increased [ 3H]thymidine incorporation by isolated canine bile duct cells. In normal dogs, the prostanoids, secretin, and glucagon increased hepatic bile flow. 10 days of ANIT feeding produced a hypercholeresis. While secretin was able to stimulate the hyperplastic biliary epithelium and increase bile flow over values produced by the hyperplastic biliary epithelium alone, neither prostaglandin E 2, prostacyclin, or glucagon appeared to stimulate the hyperplastic biliary epithelium. As ANIT produced evidence of cholestasis and hepatocellular damage, only secretin would seem to have a potential therapeutic role in increasing bile flow in cholestatic liver disorders associated with bile duct cell hyperplasia.

Inflammatory properties of bile from dogs with pigment gallstones

Gallbladder inflammation and mucus hypersecretion are prominent features of cholesterol and pigment gallstones in humans and animals. The factors leading to inflammation and mucus hypersecretion are poorly understood. These studies examine the inflammatory potential of bile from dogs with pigment gallstones. Dogs fed a methionine-deficient diet that produces pigment gallstones by 6 weeks were compared to normal dogs. Mucus layer thickness, myeloperoxidase activity, and interleukin-1-like activities were measured in canine gallbladder. The inflammatory potential of canine bile was determined by measuring mucus layer thickness, sodium absorption, myeloperoxidase activity and interleukin-1-like activity in guinea pig gallbladders exposed to normal and lithogenic canine bile for 4 hours. Mean mucus layer thickness, myeloperoxidase, and interleukin-1 activity were significantly greater in canine gallbladders containing pigment gallstones. Bile from dogs with pigment gallstones markedly increased mucus layer thickness, myeloperoxidase activity, and interleukin-1 activity and decreased sodium absorption in normal guinea pig gallbladder. These effects were not eliminated by centrifuging bile to remove crystals and gallstones. Canine bile from dogs with pigment gallstones contains soluble factors capable of causing inflammation in the gallbladder wall.

Chronic bile diversion to the urinary bladder enhances cholecystokinin release and suppresses gastric inhibitory polypeptide release in dogs.

The role of intestinal luminal bile on cholecystokinin (CCK) and gastric inhibitory polypeptide (GIP) release was examined in dogs with chronic external bile diversion. In 6 mongrel dogs, cholecysto-jejuno-cystostomy (C-J-C) was performed using a small segment of the middle small intestine interposed between the gallbladder and urinary bladder with section of the common bile duct. Butter dissolved in 30 ml of lukewarm water was orally ingested before surgery, and same amount of butter solution with or without graded volumes of canine bile were orally ingested between four to six weeks after C-J-C surgery. Increases in both plasma triglyceride and GIP levels after butter ingestion were almost completely abolished by C-J-C, but they were restored by oral bile feeding in a volume-dependent manner. Both basal and fat-stimulated CCK release were enhanced significantly by C-J-C, and oral bile feeding inhibited fat-stimulated CCK release by bile in a dose dependent manner. These data suggest that intraluminal bile regulates basal and fat-stimulated CCK release and fat-stimulated GIP release in dogs.

Does hyperamylasemia in choledochal cyst indicate true pancreatitis? An experimental study.

Patients with choledochal cyst often have repeated attacks of abdominal pain accompanied by hyperamylasemia, and they may be diagnosed as having acute pancreatitis. However, the attacks generally tend to subside in a short period by conservative treatment, and evidence of pancreatitis is rarely observed at the time of surgery. Choledochal cyst is commonly associated with pancreatobiliary maljunction, and high concentrations of pancreatic enzymes in bile are usually observed. When the bile duct pressure increases due to obstructive cholangitis, pancreatic enzymes in bile may regurgitate into the blood stream. Cholangiovenous reflux of amylase might cause hyperamylasemia. In order to investigate the mechanism of hyperamylasemia by cholangiovenous reflux, canine pancreatic juice or bile from a patient with choledochal cyst was injected into the obstructed common bile duct in dogs. The pancreatic enzymes in bile could readily enter into the blood stream at the pressure level of 15 mmHg or more in the bile duct. The peak amylase level in the thoracic lymph was observed to be more than 4 times higher than that in the blood serum, and the lymph flow during 30 minutes increased significantly from 8.1 to 20.4 ml at the bile duct pressure level of 20 mmHg. The reflux of amylase in bile into the blood stream via both the hepatic vein and thoracic duct might result in hyperamylasemia in the patients with choledochal cyst.

Comparison of covered versus uncovered wire mesh stents in the canine biliary tract

Self-expanding wire mesh stents have been developed for endoscopic placement across malignant biliary strictures, but tumor ingrowth may limit the usefulness of open mesh stents. We reasoned that coating the wire mesh might prevent tumor ingrowth. Tissue response to covered and uncovered stents was compared in dogs. Stents were surgically placed in the bile ducts of 22 mongrel dogs through the sphincter of Oddi. Either a silicone-covered stent or an uncovered stent was inserted. Liver function test values remained normal throughout a 1- or 3-month study. Necropsy revealed that all ducts were unobstructed. Bile duct histologic examination revealed mild-to-moderate cellular infiltration in all animals. Mucosal hyperplasia was more marked in the animals with uncovered stents and the bare wires became deeply embedded in bile duct epithelium, whereas the wires of covered stents did not. We conclude that covered stents are well tolerated by the canine bile duct. These results suggest that such stents may be removable, making self-expanding metal stents an appropriate treatment for both benign and malignant biliary strictures.

Comparison of covered versus uncovered wire mesh stents in the canine biliary tract

Self-expanding wire mesh stents have been developed for endoscopic placement across malignant biliary strictures, but tumor ingrowth may limit the usefulness of open mesh stents. We reasoned that coating the wire mesh might prevent tumor ingrowth. Tissue response to covered and uncovered stents was compared in dogs. Stents were surgically placed in the bile ducts of 22 mongrel dogs through the sphincter of Oddi. Either a silicone-covered stent or an uncovered stent was inserted. Liver function test values remained normal throughout a 1- or 3-month study. Necropsy revealed that all ducts were unobstructed. Bile duct histologic examination revealed mild-to-moderate cellular infiltration in all animals. Mucosal hyperplasia was more marked in the animals with uncovered stents and the bare wires became deeply embedded in bile duct epithelium, whereas the wires of covered stents did not. We conclude that covered stents are well tolerated by the canine bile duct. These results suggest that such stents may be removable, making self-expanding metal stents an appropriate treatment for both benign and malignant biliary strictures. (Gastrointest Endosc 1994; 40:17-21.)

The effect of FK506 on canine bile flow.

While the primary immunosuppressive agents utilized in solid-organ transplantation are steroids and cyclosporine, the more recently introduced agent FK506 is assuming a progressively more important role as an immunosuppressant, particularly in liver transplantation. While the effect of cyclosporine on rat and canine bile flow has been well evaluated, no information is available regarding the effect of FK506 on bile flow. Dogs with chronic biliary fistulas were utilized, enabling unanesthetized animals to be studied. FK506 was administered intravenously in varying doses, and bile volume, bile salts, and bile electrolytes were measured. FK506 produced dose-related increases in bile volume and bile chloride concentration and output, with 8 micrograms/kg-1hr-1 being the maximal dose. To ascertain that the response was not osmotic in association with FK506 secretion in bile, 500 micrograms/kg-1hr-1 FK506 was administered, which did not produce a choleresis significantly greater than 8 micrograms/kg-1hr-1. FK506 was subsequently administered orally in daily dose of 0.15 mg/kg-1 for two weeks. Oral FK506 did not consistently increase bile flow, as evaluated by a bile salt dose-response curve (9, 18, 36 mumol/min sodium taurocholate) but did significantly increase bile chloride secretion. Two weeks of oral administration of FK506 in therapeutic doses did not significantly alter serum bile salt concentrations. The results of this study indicate that intravenous FK506 produces a chloride-rich choleresis in dogs.